Comparative Phytochemical Screening of Jatropha L. Species in the Niger Delta
Phytochemical studies were carried out on the leaf, stem, root and seed of four Jatropha L. species (J. curcas L., J. gossypifolia L., J. multifida L. and J. podagrica Hook) found in some parts of the Niger Delta, with a view to ascertain the inclusion of these morphologically distinct species into a genus. Qualitative and quantitative analysis of five secondary metabolites (alkaloids, tannins, saponins, flavonoids and phenols) were undertaken. The result showed that all secondary metabolites analyzed were present in all tissues studied but at different concentrations. The concentration of tannins in the leaves of the species in descending order was observed to be 7.43% for J. curcas, 6.79% for J. podagrica, 5.16% for J. multifida and 5.14% for J. gossypifolia. Concentration of saponins in the leaf and seed of the species were: J. curcas (4.89, 2.33%); J. gossypifolia (4.15, 2.37%); J. multifida (3.15, 2.44%); J. podagrica (3.15, 2.44%). Saponin concentration in the leaf and seed of J. multifida and J. podagrica was observed to be the same suggesting close affinity. Generally, tannins were the most abundant in plant parts, followed by saponins, alkaloids, flavonoids and then phenols. Results obtained confirm the relatedness of these species and spotlight these important phytochemicals in the species. However, variations observed in their concentrations confer individuality on the species.
Received: January 24, 2011;
Accepted: March 08, 2011;
Published: July 02, 2011
Jatropha L. (Euphorbiaceae) is a very diverse subtropical and tropical
genus which includes succulents, caudiciform species, herbaceous perennials
and woody trees. Four of the eight species recorded in West Tropical Africa
and described by Hutchinson and Dalziel (1958) occur in
the Niger Delta. These are J. curcas L., J. gossypifolia L., J.
multifida L. and J. podagrica Hook. Ratha and
Paramathma (2009) described 12 species of Jatropha in India, using
morphological characters. A number of Jatropha species especially
J. curcas, yield oils of biofuel value (Agarwal and
Agarwal, 2007; Fairless, 2007; Akbar
et al., 2009), a fact which has brought much attention to the genus
particularly in India. The oil is also used for manufacture of candles and soap
and in cosmetics industry. J. curcas can be used to prevent and/or control
erosion, to reclaim land, grown as a live fence especially to contain or exclude
farm animals and be planted as a commercial crop (Kumar
and Sharma, 2008; Openshaw, 2000; Martinez-Herrera
et al., 2006). These characteristics along with its versatility make
Jatropha of vital importance to developing countries. Apart from the
earlier morphological description of on the West African species and comparison
based on epidermal features (Olowokudejo, 1993), much
is not known on the common Jatropha species in the Niger Delta.
Secondary metabolites are phytochemicals produced as by-products of primary
metabolism (Bako and Aguh, 2007) and are less widespread
in plants. It is of course this restricted occurrence among plants that renders
them valuable and useful in taxonomic delimitation of species. The groups of
compounds mostly utilized for this purpose include alkaloids, phenolics, glucosinolates,
amino acids, terpenoids, oils and waxes, carbohydrates and crystals. Recent
investigations and reports of the usefulness of these compounds either for medicinal/therapeutic
purposes or taxonomic elucidation of species include Edeoga
et al. (2005), Hassan et al. (2007),
Ibrahim et al. (2007), Mallikharjunah
et al. (2007), Irshad et al. (2010),
Nyananyo et al. (2010), Rasool
et al. (2010) and Ganesh and Vennila (2011).
The only phytochemical report known to the authors on Jatropha species
occurring in the Niger Delta is that of Burkill (1994).
This simply reported the occurrence of tannins in the leaves of J. gossypifolia
and saponin in the leaf of J. multifida. In view of the importance of
these species as highlighted and the seemingly lack of systematic information
on the species in the Niger Delta, the present study subjected the species to
qualitative and quantitative phytochemical screening. This is with a view to
verify the presence or absence of these classes of secondary metabolites in
the species as well as ascertain their relatedness or not. The study is a precursor
aimed at stimulating interest on members of this genus.
MATERIALS AND METHODS
The plant materials viz., leaf, stem, seed and root of the Jatropha species
were collected from different localities in the Niger Delta between September
2009 and March 2010. The research was conducted from March to October 2010.
The plants were identified with the help of The Flora of West Tropical Africa
(Hutchinson and Dalziel, 1958) and are deposited in the
Herbarium, Department of Plant Science and Biotechnology, University of Port
Qualitative and quantitative determination of secondary metabolites in Jatropha
species: Extracts of samples of leaf, stem, seed and root from each species
were subjected to preliminary screening for the presence of secondary metabolites
(alkaloids, tannins, flavonoids, phenols and saponins) using standard procedures
(Trease and Evans, 1989) with some modifications. The
test procedure for each compound is presented below.
Alkaloids: Five milliliter of 2% HCl was added to 2 mL of each plant
extract in a test tube placed on a steam bath and warmed. It was filtered and
divided into two parts for the following tests:
||A few drops of Wagners Reagent (Potassium-iodine solution)
were added to one part of the filtrate in a test tube. A reddish brown precipitate
||A few drops of Meyers Reagent (Potassium mercuric iodine solution)
were added to the other part of the filtrate in a test tube. A cream coloured
precipitate was observed
In both cases, the formation of precipitation indicated the presence of alkaloids.
Tannins: Five milliliter of the extract was treated with 2 mL of HCl and boiled for 5 min. The presence of red precipitate confirmed the presence of tannins.
Flavonoids: One milliliter of the extract was treated with 1 mL of dilute NaOH. The presence of a cloudy precipitate confirmed the presence of flavonoids.
Saponins: One milliliter of the extract was added to 4 mL of distilled water and shaken. A stable frothing or foaming indicated the presence of saponins.
Phenols: One milliliter of the extract was added to 1 mL of 10% FeCl2 and mixed together. The presence of blue precipitate confirmed the presence of phenols.
After preliminary analysis to determine presence of these phytochemicals, the
samples were further subjected to quantitative analysis to determine the percentage
of each of these secondary metabolites in each plant part of each species. The
following procedures were adopted:
||Quantitative analysis of alkaloids by the gravimetric method
of Harborne (1993)
Percentage alkaloids were computed as follows:
|| Weight of residue.
Tannins were computed thus:
||Flavonoid determination by the hydrolysis gravimetric method
of Harborne (1993)
Percentage Flavonoid was calculated by:
|| Weight of residue.
||Phenol analysis using the follin- ciocaltean colorimetric
method of Harborne (1993):
||Weight of sample analyzed
||Concentration of standard in mg/ml
||Total filtrate volume
||Volume of filtrate analyzed
||Dilution factor where applicable
||Saponin in plant sample was determined by the combined solvent
Saponin concentration was computed thus:
|| Average weight of residue.
The experiments were performed in triplicates (or three times) for each parameter
and the resultant percentages were summed up to deduce the average percentage
for comparison of means for each parameter.
Alkaloids Qualitative assessment (Table 1) revealed that alkaloids were observed to be deeply present (++) in the leaves and present (+) in the root and stem of all four species. However, the quantitative analysis results (Table 2) show alkaloid distribution in the different plants to be in the following decreasing order: leaf (J. curcas 4.54%; J. multifida 2.85%; J. gossypifolia 2.81%; J. podagrica 0.33%), stem (J. gossypifolia 2.16%; J. curcas 1.94%; J. multifida 1.72%; J. podagrica 0.15%), seed (J. curcas 2.85%; J. multifida 2.63%; J. gossypifolia 2.36%; J. podagrica 0.18%) and root (J. curcas 1.83%; J. multifida 1.68%; J. gossypifolia 1.60%; J. podagrica 0.26%). A similar order was observed in the leaf and root.
Tannins: Tannin was present (+) in the stem of the four species investigated. It was very deeply present (+++) in the leaf of J. curcas and J. multifida and deeply present (++) in the seed of J. curcas and J. gossypifolia and root of J. multifida (Table 1). A decreasing order of the observed results of quantitative analysis of tannin concentration in the leaves and seeds of the species follows thus: leaf (J. curcas 7.43%; J. podagrica 6.79%; J. multifida 5.16%; J. gossypifolia 5.14%), seed (J. curcas 4.23%; J. multifida and J. podagrica 3.89% each; J. gossypifolia 3.52%). Also, the same tannin concentration of 2.73% (J. gossypifolia and J. podagrica) and 1.21% (J. multifida and J. podagrica) were recorded in the root and stem of the species respectively (Table 2).
Flavonoids: Flavonoids were observed to be very deeply present (+++) in the leaves of J. curcas and J. multifida and deeply present (++) in that of J. gossypifolia. They were observed to be present (+) in all other plant parts of the species (Table 1). The highest flavonoid concentration of 3.25% was observed in the leaf of J. podagrica (Table 2). Flavonoid was 2.76% in J. curcas leaf, 2.41% in J. gossypifolia and 2.18% in J. multifida. The same concentration of 2.14% (seed) and 1.28% (stem) were observed in J. multifida and J. podagrica. The least flavonoid concentration of 1.20% was observed stem of J. gossypifolia.
|| Preliminary (qualitative) screening of secondary metabolites
in Jatropha species
|+: Present, ++: Deeply present, +++: Very deeply present
|| Concentration of secondary metabolites present in Jatropha
Saponins: The saponins were deeply present (++) in the leaf of the four species and seed of J. podagrica. They were observed to be present (+) in all other plant parts of the species. A descending order of the concentration of saponins in the leaf and stem of the species as revealed from the quantitative analysis follows thus: leaf (J. curcas 4.89%; J. gossypifolia 4.15%; J. multifida and J. podagrica 3.15% each), stem (J. podagrica 2.77%; J. multifida 2.73%; J. curcas 2.27%; J. gossypifolia 2.18%). The least saponin concentration of 2.16% was observed in the root of J. multifida while the highest concentration of 4.89% was in the leaf of J. curcas.
Phenols: Phenols were generally low qualitatively and quantitatively.
They were observed to be deeply present (++) only in leaves of J. curcas
and J. gossypifolia and present in all other plant parts investigated
in the four species. The highest and least phenol concentrations of 0.59 and
0.13% were recorded in the stems of J. curcas and J. gossypifolia,
respectively. While the seed of J. podagrica, J. multifida and
J. gossypifolia were observed to have the same phenol concentration of
0.18%, the leaves of J. multifida and J. gossypifolia had 0.16%.
The results of the comparative qualitative and quantitative phytochemical analysis
of secondary metabolites in the leaf, root, seed and stem of Jatropha
species in the Niger Delta are presented in Tables 1 and 2.
Burkill (1994) reported the presence of tannins and saponins
in the leaf of J. gossypifolia and J. multifida respectively which
is confirmed by this study. The qualitative screening from the presence study
revealed the presence of alkaloids, tannins, saponins, flavonoids and phenols
in all plant parts investigated but to varying intensities/qualities as shown
in Table 1. Preliminary qualitative test according to Mallikharjunah
et al. (2007) is useful in the detection of bioactive principles
and subsequently may lead to drug discovery and development. Qualitative phytochemical
screening of two species of Avicennia (Ganesh and Vennila,
2011) and three cucurbits (Irshad et al., 2010)
revealed the presence of alkaloids, flavonoids, tannins, saponins and phenols
in a manner suggesting close relationship of the species studied. Similar results
(Table 1) were obtained in this research work which confirm
the relatedness of the investigated Jatropha species as well as reveal
their potentials in the drug industry. Specifically, the qualitative screening
revealed that tannins were very deeply present in the leaves and stems of
J. curcas and J. multifida and deeply present in the roots of
J. curcas, J. gossypifolia and J. multifida. Likewise, alkaloids
were deeply present in the leaf of J. multifida, the stem of all species
except in J. multifida and the roots of J. curcas, J. gossypifolia
and J. multifida. Olawale-Abulude (2007) undertook
phytochemical screening of leaves of twenty-eight woody species from different
plants families in Nigeria and discovered the presence of tannins, alkaloids
and flavonoid in all samples. Thus these secondary metabolites seem cosmopolitan
in plants but to varying degrees and types. Their different degrees of occurrence
in plant parts confer taxonomic usefulness on them. This is similar to the findings
of Nyananyo et al. (2010) on some Niger Delta
Among the five groups of phytochemicals investigated from the leaf, root, seed
and stem of the species, tannins were found to be the most abundant followed
by saponins and alkaloids. Phenols were the lowest in concentration and together
with flavonoids varied across the different plant parts. Specifically, tannins
were concentrated to 7.43% in the leaves of J. curcas with corresponding
concentration of 0.38% by phenols. Similarly, saponin (the most abundant phytochemical
in the root of the species) with concentration of 3.62% (the highest in all
seed samples) in the root of J. curcas had corresponding phenol concentration
of 0.20%. A similar sequence with tannins being the most abundant phytochemical
in plant parts, followed by saponins, alkaloids, flavonoids and then phenols
as seen in this study has been reported by Mallikharjunah
et al. (2007). Other recent investigation showcasing presence, distribution
in plant parts and importance of these phytochemicals similar with the results
of this investigation include Ogunkunle and Ladejobi (2006),
Sunita and Abhishek (2008), Ferreira
et al. (2009), Kumar et al. (2009)
and Ganesh and Vennila (2011). It is noteworthy that while
all these phytochemicals are present in the different Jatropha species
connoting taxonomic affinity, the differences in their concentrations uniquely
confers individualism on each species and thus support their being treated as
taxonomic species. This agrees with Irshad et al.
(2010) where phytochemical results established closer relationship between
Lagenaria siceraria and Luffa cylindrical than Cucumis maxima.
This study reveals the presence of the subject secondary metabolites in the
different Jatropha species to varying concentrations which is taxonomically
useful; it also brings to bare the fact that the species are potential sources
of these important phytochemicals. For instance, flavonoids are one of the most
popular secondary metabolites possessing a variety of biological activities
at nontoxic concentrations (Irshad et al., 2010).
Dietary flavonoids are noted to play effective roles in cancer prevention (Ren
et al., 2003; Aggarwal and Shishodia, 2006).
Flavonoids together with the other secondary metabolites identified in the Jatropha
species have been severally reported in other plants to show curative activity
against diverse pathogens, used traditionally as analgesic, antimicrobial and
soothing herbs (Hassan et al., 2004; Faruq
et al., 2004; Olafimihan, 2004; Singh
et al., 2009; Thirunavukkarasu et al.,
2010; Ganesh and Vennila, 2011).
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